Le K D, Gilkes N R, Kilburn D G, Miller R C, Saddler J N, Warren R A
Department of Microbiology, University of British Columbia, Vancouver, Canada.
Enzyme Microb Technol. 1994 Jun;16(6):496-500. doi: 10.1016/0141-0229(94)90020-5.
A fusion protein, Sta-CBDCex, which comprises streptavidin with a cellulose-binding domain (CBDCex) fused to its C terminus, was produced in the cytoplasm of Escherichia coli, where it formed inclusion bodies. Renatured Sta-CBDCex, recovered from the inclusion bodies, adsorbed to Avicel, a microcrystalline cellulose. The cellulose-bound Sta-CBDCex in turn bound biotinylated alkaline phosphatase or biotinylated beta-glucosidase. The immobilized beta-glucosidase remained fully active during 2 weeks of continuous column operation at 50 degrees C.
一种融合蛋白Sta-CBDCex，其C末端融合了具有纤维素结合结构域（CBDCex）的链霉亲和素，在大肠杆菌细胞质中产生，在那里它形成了包涵体。从包涵体中回收的复性Sta-CBDCex吸附到微晶纤维素Avicel上。结合在纤维素上的Sta-CBDCex进而结合生物素化的碱性磷酸酶或生物素化的β-葡萄糖苷酶。固定化的β-葡萄糖苷酶在50℃连续柱操作2周期间保持完全活性。
